A Tale of Two Biomarkers: Untargeted 1H NMR Metabolomic Fingerprinting of BHBA and NEFA in Early Lactation Dairy Cows

Abstract

Disorders of energy metabolism, which can result from a failure to adapt to the period of negative energy balance immediately after calving, have significant negative effects on the health, welfare and profitability of dairy cows. The most common biomarkers of energy balance in dairy cows are β-hydroxybutyrate (BHBA) and non-esterified fatty acids (NEFA). While elevated concentrations of these biomarkers are associated with similar negative health and production outcomes, the phenotypic and genetic correlations between them are weak. In this study, we used an untargeted 1H NMR metabolomics approach to investigate the serum metabolomic fingerprints of BHBA and NEFA. Serum samples were collected from 298 cows in early lactation (calibration dataset N = 248, validation N = 50). Metabolomic fingerprinting was done by regressing 1H NMR spectra against BHBA and NEFA concentrations (determined using colorimetric assays) using orthogonal partial least squares regression. Prediction accuracies were high for BHBA models, and moderately high for NEFA models (R2 of external validation of 0.88 and 0.75, respectively). We identified 16 metabolites that were significantly (variable importance of projection score > 1) correlated with the concentration of one or both biomarkers. These metabolites were primarily intermediates of energy, phospholipid, and/or methyl donor metabolism. Of the significant metabolites identified; (1) two (acetate and creatine) were positively correlated with BHBA but negatively correlated with NEFA, (2) nine had similar associations with both BHBA and NEFA, (3) two were correlated with only BHBA concentration, and (4) three were only correlated with NEFA concentration. Overall, our results suggest that BHBA and NEFA are indicative of similar metabolic states in clinically healthy animals, but that several significant metabolic differences exist that help to explain the weak correlations between them. We also identified several metabolites that may be useful intermediate phenotypes in genomic selection for improved metabolic health.

Metabolomics comparison of serum and urine in dairy cattle using proton nuclear magnetic resonance spectroscopy

“…According to Xu et al [12], changes in the concentrations of acetone, arginine, BHBA, glucose, glycine, kynurenine, and panthothenate in the plasma may be an indicator of NEB. Further, as reported by Luke et al [15], an increase in the BHBA concentrations in early lactation dairy cattle was positively correlated with acetate, betaine, creatine, glycine, and phosphocholine, and negatively correlated with alanine, dimethyl sulfone, glucose, lactate, and valine in the serum. All metabolites associated with NEB were measured in this study, except for kynurenine; further, the metabolites associated with increased BHBA concentration in dairy cattle during early lactation were also measured in this study, except for dimethyl sulfone and phosphocholine.…”

“…of ruminants has been actively conducted worldwide. Metabolomics research using ruminant serum and plasma have been conducted to investigate the relationship between negative energy balance (NEB) and early lactation in dairy cows [ 12 ], feed efficiency in black angus steers [ 13 ], heat stress (HS) in beef cattle breeds [ 14 ], and in search of biomarkers of ketosis and bovine respiratory diseases (BRD) [ 15 , 16 ]. Research on metabolomics using ruminant urine has focused on urea metabolism and recycling [ 17 ], postpartum diseases such as metritis and lameness in dairy cows [ 18 , 19 ], HS in beef cattle breeds [ 14 ], and nitrogen efficiency in dairy cows [ 20 ].…”

A Comparison of Rice Husks and Peanut Shells as Bedding Materials on Dairy Cows’ Preferences, Behaviour, and Health

“…Adequate energetic balance is important for maintaining both cow welfare and adequate production levels. Serum BHB and NEFA concentrations are used as indicators of energy balance and hyperketonemia in cows [42,43]. Ketosis can be diagnosed by a BHB serum concentration of ≥1.2 mmol/L [32,44].…”

The Dynamic Transcription Profiles of Proliferating Bovine Ovarian Granulosa When Exposed to Increased Levels of β-Hydroxybutyric Acid

“…This is due to an insufficiency in the dry matter intake (DMI) and high levels of lactation. If cows cannot adapt to NEB in a timely manner, they are prone to metabolic disorders and produce excess ketone bodies (KBs), which may ultimately develop ketosis (1)(2)(3)(4). Ketosis is characterized by an increase of KBs in the blood, with βhydroxybutyric acid (BHBA) being approximately 70% of the KBs (5).…”